Background/Aims: Liver transplantation (LT) following total hepatectomy is a life-saving treatment for hepatocellular carcinoma (HCC). The HCC recurrence after LT hinders the effectiveness of the procedure. The objective of this study is to develop a pre-operative risk stratification model based on a liquid biopsy. Methods: We conducted a comprehensive multi-omics study of 260 HCC patients from three centers, including clinical data, low-coverage whole-genome sequencing of cell-free DNA (cfDNA) from plasma, as well as whole-exome, single-nucleus RNA, and spatial transcriptomics from matched tumor and non-tumor tissues. Results: We identified cfDNA-derived copy number alteration (CNA) signatures associated with post-transplant recurrence. By integrating cfDNA-derived CNA profiles with single-cell transcriptomic data, we traced recurrence-associated cfDNA to a distinct subpopulation of malignant cells within the primary tumor. These cells were embedded in a pro-metastatic microenvironment of specialized endothelial subtypes and cancer-associated fibroblasts. Notably, most recurrence-associated lesions were detectable in cfDNA prior to liver transplantation (LT). Building on these insights, we developed the ZJU Criteria based on CNA fragments and tumor markers, a pre-LT risk prediction tool that integrates conventional clinical factors with cfDNA-derived CNA signatures, and validated it using internal and independent external cohorts. Conclusion Our findings suggest that post-transplant recurrence commonly originates from advanced subclones that emerge late during tumor evolution. The ZJU Criteria provides an accurate, non-invasive strategy that significantly improves pre-LT risk stratification and clinical decision-making for patients with HCC.

Objective To investigate the expression characteristics, prognostic value, and correlation with immune infiltration of vasoactive intestinal peptide receptor 2 (VIPR2) in esophageal adenocarcinoma (ESAD). Methods Transcriptome data of 80 ESAD and 10 normal tissues were retrieved from the TCGA database. Immune-related differential genes were identified by integrating the ImmPort database. Cox regression analysis was applied to evaluate the prognostic significance of VIPR2. GSVA and ssGSEA were employed to assess the association between VIPR2 and immune-cell infiltration. Functional enrichment was performed using GO, KEGG, and GSEA. Results VIPR2 expression was significantly downregulated in ESAD tissues (P<0.05). High VIPR2 expression was associated with prolonged overall survival (HR=0.246, P=0.047), serving as an independent prognostic factor. Fifteen immune-cell types, including CD8⁺ T cells, NK cells, and dendritic cells, showed significantly higher infiltration in the VIPR2-high group, whereas Th17-cell abundance was decreased (P<0.05). Enrichment analyses revealed that VIPR2 participated in multiple tumor-immune pathways and was positively correlated with PD-1 and CTLA-4 expression (P<0.01). Conclusion VIPR2 may serve as a potential biomarker in the prognostic assessment and could modulate the tumor-immune microenvironment in ESAD.

Objective To examine the changing prevalence and antimicrobial resistance profiles of Burkholderia cepacia in 52 hospitals across China from 2015 to 2021.Methods A total of 9 261 strains of B.cepacia were collected from 52 hospitals between January 1,2015 and December 31,2021.Antimicrobial susceptibility of the strains was tested using Kirby-Bauer method or automated antimicrobial susceptibility testing systems according to a unified protocol.The results were interpreted according to the breakpoints released in the Clinical & Laboratory Standards Institute(CLSI)guidelines(2023 edition).Results A total of 9 261 strains of B.cepacia were isolated from all age groups,especially elderly patients.The proportion was 11.1％(1 032 strains)in children,significantly lower than the proportion in adults.About half(46.5％,4 310/9 261)of the strains were isolated from patients at least 60 years old and 42.3％(3 919/9 261)of the strains were isolated from young adults.Most isolates(71.1％)were isolated from sputum and respiratory secretions,followed by urine(10.7％)and blood samples(8.1％).B.cepacia isolates were highly susceptible to the five antimicrobial agents recommended in the CLSI M100 document(33rd edition,2023).B.cepacia isolates showed relatively higher resistance rates to meropenem and levofloxacin.However,the resistance rates to ceftazidime,trimethoprim-sulfamethoxazole,and minocycline remained below 8.1％.The percentage of B.cepacia strains resistant to levofloxacin was the highest compared to other antibiotics in any of the three age groups(from 12.4％in the patients＜18 years old to 20.6％in the patients aged 60 years or older).Conclusions B.cepacia is one of the clinically important non-fermenting gram-negative bacteria.Accurate and timely reporting of antimicrobial susceptibility test results and ongoing antimicrobial resistance surveillance are helpful for rational prescription of antimicrobial agents and proper prevention and control of nosocomial infections.

Objective To analyze the risk factors of fluconazole resistance in Candida tropicalis(C.tropicalis)urinary tract infection(UTI),and evaluate the efficacy of different treatment regimens.Methods Patients with C.tropicalis UTI at Xiangya Hospital of Central South University from January 2021 to December 2023 were in-cluded for single center retrospective study.The minimum inhibitory concentration(MIC)of fluconazole was deter-mined by microbroth dilution.Patients were divided into a fluconazole-resistant group and a fluconazole-sensitive group based on fluconazole resistance.Risk factors for fluconazole resistance were analyzed based on clinical data,and therapeutic efficacy in patients in fluconazole-resistant group was analyzed.Results A total of 198 patients were included in the study.133(67.2％)C.tropicalis strains were detected to be sensitive to fluconazole,while 65(32.8％)strains were resistant,and 63.1％(n=41)had MIC values ≥128 μg/mL.Compared with fluconazole-sensitive group,fluconazole-resistant group had a higher proportion of pulmonary infection(P=0.019).Pulmonary infection(OR=3.282)was a risk factor for fluconazole resistance in C.tropicalis UTI,while urinary system ob-struction(OR=0.269)was a protective factor for fluconazole resistance in C.tropicalis UTI.There was no statis-tically significant difference in the usage rate of different antimicrobial agent types between the two groups(all P＞0.05).The therapeutic efficacy analysis showed that the effective rates of treatment with fluconazole dosage regi-mens of ≤200 mg/d,≥400 mg/d,and fluconazole monotherapy against fluconazole-resistant strains were 66.7％(6/9),83.3％(5/6),and 100％(6/6),respectively.For patients treated with monotherapy using other drugs or with multidrug sequential treatment regimens,the treatment effective rate was 60.0％(3/5).The proportion of pa-tients in the effective treatment group who removed their urinary catheters after detecting C.tropicalis was higher than that in the ineffective treatment group(P＜0.001).Conclusion The fluconazole resistance of C.tropicalis is related to urinary tract obstruction and concurrent pulmonary infection.When treating UTI caused by fluconazole-resistant strains,the catheter should be removed as early as possible.In addition to increasing the dosage of flucon-azole,other antifungal drugs such as flucytosine alone or sequential treatment with multiple drugs can also be con-sidered.

Objective To summarize the changing prevalence of carbapenem resistance in Enterobacterales based on the data of CHINET Antimicrobial Resistance Surveillance Program from 2015 to 2021 for improving antimicrobial treatment in clinical practice.Methods Antimicrobial susceptibility testing was performed using a commercial automated susceptibility testing system according to the unified CHINET protocol.The results were interpreted according to the breakpoints of the Clinical & Laboratory Standards Institute(CLSI)M100 31st ed in 2021.Results Over the seven-year period(2015-2021),the overall prevalence of carbapenem-resistant Enterobacterales(CRE)was 9.43％(62 342/661 235).The prevalence of CRE strains in Klebsiella pneumoniae,Citrobacter freundii,and Enterobacter cloacae was 22.38％,9.73％,and 8.47％,respectively.The prevalence of CRE strains in Escherichia coli was 1.99％.A few CRE strains were also identified in Salmonella and Shigella.The CRE strains were mainly isolated from respiratory specimens(44.23±2.80)％,followed by blood(20.88±3.40)％and urine(18.40±3.45)％.Intensive care units(ICUs)were the major source of the CRE strains(27.43±5.20)％.CRE strains were resistant to all the β-lactam antibiotics tested and most non-β-lactam antimicrobial agents.The CRE strains were relatively susceptible to tigecycline and polymyxins with low resistance rates.Conclusions The prevalence of CRE strains was increasing from 2015 to 2021.CRE strains were highly resistant to most of the antibacterial drugs used in clinical practice.Clinicians should prescribe antimicrobial agents rationally.Hospitals should strengthen antibiotic stewardship in key clinical settings such as ICUs,and take effective infection control measures to curb CRE outbreak and epidemic in hospitals.

Objective To characterize the changing species distribution and antibiotic resistance profiles of respiratory isolates in hospitals participating in the CHINET Antimicrobial Resistance Surveillance Program from 2015 to 2021.Methods Commercial automated antimicrobial susceptibility testing systems and disk diffusion method were used to test the susceptibility of respiratory bacterial isolates to antimicrobial agents following the standardized technical protocol established by the CHINET program.Results A total of 589 746 respiratory isolates were collected from 2015 to 2021.Overall,82.6％of the isolates were Gram-negative bacteria and 17.4％were Gram-positive bacteria.The bacterial isolates from outpatients and inpatients accounted for(6.0±0.9)％and(94.0±0.1)％,respectively.The top microorganisms were Klebsiella spp.,Acinetobacter spp.,Pseudomonas aeruginosa,Staphylococcus aureus,Haemophilus spp.,Stenotrophomonas maltophilia,Escherichia coli,and Streptococcus pneumoniae.Each microorganism was isolated from significantly more males than from females(P＜0.05).The overall prevalence of methicillin-resistant S.aureus(MRSA)was 39.9％.The prevalence of penicillin-resistant S.pneumoniae was 1.4％.The prevalence of extended-spectrum β-lactamase(ESBL)-producing E.coli and K.pneumoniae was 67.8％and 41.3％,respectively.The overall prevalence of carbapenem-resistant E.coli,K.pneumoniae,Enterobacter cloacae,Pseudomonas aeruginosa,and Acinetobacter baumannii was 3.7％,20.8％,9.4％,29.8％,and 73.3％,respectively.The prevalence of β-lactamase was 96.1％in Moraxella catarrhalis and 60.0％in Haemophilus influenzae.The H.influenzae isolates from children(＜18 years)showed significantly higher resistance rates to β-lactam antibiotics than the isolates from adults(P＜0.05).Conclusions Gram-negative bacteria are still predominant in respiratory isolates associated with serious antibiotic resistance.Antimicrobial resistance surveillance should be strengthened in clinical practice to support accurate etiological diagnosis and appropriate antimicrobial therapy based on antimicrobial susceptibility testing results.

Eightly-four novel thioheterocyclic nucleoside derivatives were designed, synthesized, and evaluated for antitumor activity in vitro and in vivo. Most of the compounds inhibited the growth of HCT116 and HeLa cancer cells in vitro, among them 33a and 36b exhibited potent activity against HCT116 cells (IC₅₀ = 0.27 and 0.49 μmol/L, respectively). Both compounds 33a and 36b inhibited cell metastasis, arrested the cell cycle in the G₂/M phase, and induced apoptosis in vitro. Mechanistic studies revealed that 33a and 36b increased ROS levels, led to DNA damage, ER stress, and mitochondrial dysfunction, and inhibited autophagy in HCT116 cells. Biological information analysis, RNA-sequencing, Gene Set Enrichment Analysis (GSEA), drug affinity responsive target stability (DARTS) assay, cellular thermal shift assay (CETSA), and SPR experiments identified that compounds 33a and 36b showed antitumor activity by suppressing the c-MYC pathway. c-MYC silencing assays indicated that c-MYC proteins participated in 33a-mediated anticancer activities in HCT116 cells. More importantly, compound 33a presented favorable pharmacokinetic properties in mice (T _1/2 = 6.8 h) and showed significant antitumor efficacy in vivo without obvious toxicity, showing promising potential for further clinical development.

Isoflavones which mainly distributed in leguminous plants have plenty of health benefits.Isoflavone synthase(IFS)is a membrane-associated cytochrome P450 enzyme(CYP450)which carries out the unique aryl-ring migration and hydroxylation.So far,few crystal structures of plant P450s have been obtained.We determined the crystal structure of IFS from Medicago truncatula at 1.9 ? by MAD method using a selenomethionine substituted crystal and conducted molecular docking and mutagenesis study.The structure of IFS complexed with imidazole exhibits the helix Ⅰa-loop-helix Ⅰβ motif which cor-responds to helix Ⅰ of other P450s.Compared with structures of common P450s,IFS/imidazole structure contains an extra domain,i.e.,the γ-domain.The structure reveals a homodimer in which the γ-domain of one molecule interacts with the β-domain of another.The plane of heme group makes an angle of ap-proximately 40° with the helix Ⅰa-loop-helix Ⅰβ motif.Molecular docking combined with mutagenesis study suggested that Trp-128 and Asp-300 might play important roles in substrate binding and recogni-tion.Phe-301,Ser-303 and Gly-305 from the helix Ⅰa-loop-helix Ⅰβ motif may play important roles in the aryl-ring migration.These novel structural features reveal insights into the unique reaction mechanism of IFS and provide a basis for engineering IFS in leguminous crops for health purpose.

Objective:To investigate the surgical technique and initial outcomes of distal derotational femoral osteotomy (DDFO) combined with knee extension device reconstruction in adolescents with habitual patellar dislocation and severe lower limb torsional deformity.Methods:A retrospective study was conducted on 10 adolescent patients (12 knees) treated at Beijing Jishuitan Hospital and its Guizhou branch from June 2016 to June 2022. There were 6 males and 4 females with an average age of 12.0±1.5 years (range: 10.0-14.5 years) Surgical treatment included DDFO and knee extension device reconstruction (lateral retinacular release, medial retinacular plication, Roux-Goldthwait distal realignment, and MPFL reconstruction). Clinical outcomes were assessed using Lysholm scores, incidence of redislocation and complications, and imaging parameters (lateral patellofemoral angle, Insall-Salvati index, TT-TG distance, and femoral anteversion angle) preoperatively and at 1 year postoperatively.Results:All 12 knees were successfully operated on, with an average surgery time of 2.0±0.5 h (range 1.0-2.5 h), intraoperative blood loss of 47.1±17.1 ml (range 20-80 ml), and follow-up time of 46.2±18.7 months (range 24-72 months). The Lysholm knee score improved from 58.25±8.80 preoperatively to 89.17±5.32 at final follow-up ( t=-9.096, P<0.001) with significant difference. The lateral patellofemoral angle improved from -64.92±4.68 preoperatively to 6.08±2.27 at final follow-up ( t=39.178, P<0.001) with significant difference. The femoral anteversion angle decreased from 34.08±3.06 preoperatively to 14.50±2.65 at final follow-up ( t=16.916, P<0.001) with significant difference. No patellar redislocation, skin necrosis, wound infection, or limited joint mobility occurred during follow-up. Conclusion:DDFO combined with knee extension device reconstruction is an effective and safe treatment for adolescent habitual patellar dislocation with severe torsional deformity, resulting in significant clinical and radiographic improvement with low complication rates.

Objective:To evaluate the efficacy of digitally assisted design of surgical template for fibular composite tissue flap in reconstruction of segmental mandibular defect with soft tissue defect.Methods:A retrospective analysis was conducted on 30 patients who were treated at the Department of Otolaryngology Head and Neck Surgery, the 920th Hospital of the Joint Logistics Support Force of Chinese PLA from July 2020 to May 2024 for segmental mandibular defects combined with soft tissue defect. The cohort comprised 19 males and 11 females and aged 11-71 years (44.33 years±15.31 years). Pathological diagnoses of the patients were ameloblastoma (14 cases), squamous cell carcinoma (9 cases), osteomyelitis (4 cases), and odontogenic keratocyst (3 cases). Primary reconstructive surgery were performed on 21 patients after surgical resection of lesions, and 9 patients received secondary reconstructive surgery. The length of mandibular defect ranged from 75.83 mm to 111.45 mm (87.31 mm±12.00 mm), and soft tissue defects were measured at 5.0 cm×1.8 cm to 8.6 cm×2.1 cm (mean area 13.63 cm 2±2.42 cm 2). Preoperative CTA was performed to locate the perforator of peroneal artery and for design of digital surgical template. Intraoperatively, a modified fibula composite tissue flap harvesting technique was employed and that involved in: CTA-guided perforator planning, fibula osteotomy, anterior intermuscular septum exposure for identifying the peroneal artery, and a digital template-assisted fibula crafting for reconstruction of mandibule and soft tissue defect. Postoperative follow-ups were conducted at 1, 3, and 6 months, followed by quarterly reviews at outpatient clinic or via telephone interviews. Statistical analysis was performed using SPSS 27.0 software with descriptive statistical methods. Results:After surgery, all the fibula composite tissue flaps were viable and the incision wound healed well. Two patients had partial necroses at distal edge of the flaps, and they were healed after treatment. One patient had donor site infection and healed after anti-infective treatment. One month after the surgery, patients were assessed according to the recovery of face, mouth opening and occlusion, of which 25 patients (83.3%) were rated of Grade I, 4 (13.3%) of Grade Ⅱ, and 1 (3.4%) of Grade Ⅳ, with an excellent and good rates of 96.6%. The average distance of condylar movement on the affected side was 1.28 mm±0.35 mm. Postoperative follow-up lasted for 10 to 22 months, with 19.17 months±2.14 months in average. Assessment at the final follow-up was found that a total of 26 patients (86.8%) were of Grade I, 3 (10.0%) of Grade Ⅱ, and 1 (3.3%) of Grade Ⅳ and all the transferred fibula showed good alignment with the mandible over the postoperative follow-up period.Conclusion:Digital template-assisted design of fibular composite tissue flap enables a precise vascular protection, individualised osteotomy and functional restoration in reconstruction of segmental mandibular defect with soft tissue defect. It demonstrates a high clinical feasibility.